User:Matt Hartings/Notebook/Photosynthesis/2013/10/03: Difference between revisions
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|style="background-color: #EEE"|[[Image:Hartings_AU_Photosynthesis_Lab_Header.png|128px]]<span style="font-size:22px;"> Protein Re-engineering</span> | |style="background-color: #EEE"|[[Image:Hartings_AU_Photosynthesis_Lab_Header.png|128px]]<span style="font-size:22px;"> Protein Re-engineering</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==Objective== | ==Objective== | ||
Take kinetic data for Mb catalyzed o-phenylenediamine + H2O2 reaction at higher temperatures (25, 35, and 45C) | |||
==Procedure and Data== | |||
<u>Stock solutions</u> | |||
<u>sample 1</u> | |||
# Stock solutions: | |||
## Myoglobin | |||
### 2.0mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL water | |||
## OPD | |||
### 4.1mg OPD in 2.4mL water | |||
## H<sub>2</sub>O<sub>2</sub> | |||
### 0.114mL 30% H<sub>2</sub>O<sub>2</sub> in 0.887mL water | |||
#Experimental Run | |||
## Sample chamber set to 25C and to stir | |||
## N<sub>2</sub> run through the cuvette holder to reduce condensation on cuvette | |||
## 2.025mL OPD stock solution | |||
## 0.75mL H<sub>2</sub>O<sub>2</sub> stock solution | |||
### Start Data Collection | |||
### 1ms Integration | |||
### 10 scan average | |||
### Wait 1 minute after each scan | |||
### Stop after 2 hours | |||
## Files saved as: Mb_OPD_H2O2_Citrate_H20_25_SCANNUMBER.txt | |||
## 0.225mL Mb stock solution added just before the third scan | |||
## This is denoted in the data file for the third scan | |||
*Note - This run was over in a hurry. Something was off (mass of Mb or mass of OPD or both) I stopped the run early and will use the balance in 207 for the remainder of experiments. | |||
<u>sample 2</u> | |||
# Stock solutions: | |||
## Myoglobin | |||
### 1.5mg (Mb:KCl lyophilized from Citrate, 5mg:500mg) in 1.0mL water | |||
## OPD | |||
### 3.0mg OPD in 3.3mL water | |||
## H<sub>2</sub>O<sub>2</sub> | |||
### 0.114mL 30% H<sub>2</sub>O<sub>2</sub> in 0.887mL water | |||
#Experimental Run | |||
## Sample chamber set to 35C and to stir | |||
## N<sub>2</sub> run through the cuvette holder to reduce condensation on cuvette | |||
## 2.025mL OPD stock solution | |||
## 0.75mL H<sub>2</sub>O<sub>2</sub> stock solution | |||
### Start Data Collection | |||
### 1ms Integration | |||
### 10 scan average | |||
### Wait 1 minute after each scan | |||
### Stop after 2 hours | |||
## Files saved as: Mb_OPD_H2O2_Citrate_H20_35_SCANNUMBER.txt | |||
## 0.225mL Mb stock solution added just before the third scan | |||
## This is denoted in the data file for the third scan | |||
<u>sample 3</u> | |||
# Stock solutions: | |||
## Myoglobin | |||
### Used the same stock as in sample 2 | |||
## OPD | |||
### 2.5mg in 2.5mL | |||
## H<sub>2</sub>O<sub>2</sub> | |||
### 0.114mL 30% H<sub>2</sub>O<sub>2</sub> in 0.887mL water | |||
#Experimental Run | |||
## Sample chamber set to 45C and to stir | |||
## N<sub>2</sub> run through the cuvette holder to reduce condensation on cuvette | |||
## 2.025mL OPD stock solution | |||
## 0.75mL H<sub>2</sub>O<sub>2</sub> stock solution | |||
### Start Data Collection | |||
### 1ms Integration | |||
### 10 scan average | |||
### Wait 1 minute after each scan | |||
### Stop after 2 hours | |||
## Files saved as: Mb_OPD_H2O2_Citrate_H20_35_SCANNUMBER.txt [[User:Matt Hartings|Matt Hartings]] I messed up with the naming. Name should have 45 instead of 35. but file says 35 | |||
## 0.225mL Mb stock solution added just before the third scan | |||
## This is denoted in the data file for the third scan | |||
Latest revision as of 23:24, 26 September 2017
Protein Re-engineering | Main project page Previous entry Next entry |
ObjectiveTake kinetic data for Mb catalyzed o-phenylenediamine + H2O2 reaction at higher temperatures (25, 35, and 45C) Procedure and DataStock solutions sample 1
sample 2
sample 3
|